论文部分内容阅读
目的观察人类免疫缺陷病毒(HIV)感染者外周血单个核细胞(PBMC)体外培养过程中免疫学及病毒学的动态变化特点,并评价培养的细胞体系抗病毒活性。方法采集14例HIV感染者和6名健康人的PBMC,体外经多种细胞因子诱导培养,每3天观察细胞增殖情况,同时检测细胞表型、上清中细胞因子浓度和病毒载量。结果培养的健康人PBMC在(35±5)d最大增殖(61±8)倍;7例培养成功的HIV感染者PBMC在(21±6)d最大增殖(17±13)倍,另7例HIV感染者PBMC培养失败;同时发现HIV感染者PBMC体外培养最大增殖时间与其培养前外周血基础CD4/CD8比值呈明显正相关(P<0.05)。表型分析发现,培养的PBMC为优先CD8细胞增殖的异质T细胞群,主要由CD4、CD8及CD3CD56细胞组成;部分HIV感染者PBMC培养期间分泌白细胞介素(IL)1α、IL12、肿瘤坏死因子(TNF)α和IL10等细胞因子能力较高;而11/12例HIV感染者PBMC体外培养初期可扩增出大量病毒,但随着培养时间的延长,病毒载量逐渐降低甚至低于检测限。结论体外大量扩增基础CD4/CD8比值较高的HIV感染者外周血PBMC是可行的,扩增的细胞具备较强的Th1细胞免疫反应能力,为进一步开展艾滋病的免疫细胞治疗提供必要的实验数据。
Objective To observe the immunological and virological characteristics of peripheral blood mononuclear cells (PBMCs) from human immunodeficiency virus (HIV) infected patients during in vitro culture and to evaluate the antiviral activity of cultured cell lines. Methods PBMC from 14 HIV-infected and 6 healthy individuals were collected and cultured in vitro by various cytokines. Cell proliferation was observed every 3 days. Cell phenotype, cytokine concentration in supernatant and viral load were also detected. Results The PBMC of healthy people were (61 ± 8) times of that of (35 ± 5) days. The PBMC of seven HIV-infected patients were (17 ± 13) times of that of (21 ± 6) d, However, PBMC in HIV-infected patients failed to culture in vitro. At the same time, it was found that the maximum proliferative time of PBMC in HIV-infected patients was positively correlated with the ratio of CD4 / CD8 in peripheral blood before culture (P <0.05). Phenotypic analysis showed that the cultured PBMC was a heterogeneous group of T cells that preferentially proliferated CD8 cells, mainly composed of CD4, CD8 and CD3CD56 cells; some HIV-infected PBMC secreted interleukin (IL) 1α, IL12 during tumor culture, tumor necrosis (TNF) α and IL10 and other cytokines higher; and 11/12 cases of HIV-infected PBMC in early culture can amplify a large number of viruses, but with the incubation time, the viral load decreased or even lower than the detection limit. Conclusion It is feasible to extensively amplify PBMC from peripheral blood of HIV-infected patients with large CD4 / CD8 ratio in vitro. The expanded cells possess strong ability of Th1 immune response and provide necessary experimental data for further immunotherapy of AIDS. .